Yeast, beef and pork extracts counteract Clostridium difficile toxin A enterotoxicity

Clostridium difficile is responsible for a large proportion of nosocomial cases of antibiotic-associated diarrhoea and pseudomembranous colitis. The present study provides evidence that yeast, beef and pork extracts, ingredients commonly used to grow bacteria, can counteract C. difficile toxin A enterotoxicity in vitro and in vivo . In model intestinal epithelial cells the individual extracts could prevent the toxin A-induced decrease in epithelial barrier function and partially prevented actin disaggregation and cell rounding. Mice with ad libitum access to individual extracts for 1 week had almost complete reduction in toxin A-induced fluid secretion in intestinal loops. Concomitantly, the toxin A-induced expression of the essential proinflammatory mediator Cox-2 was normalized. Moreover this protective effect was also seen when mice received only two doses of extract by intragastric gavage within 1 week. These results show that yeast, beef and pork extracts have the potential to counteract the intestinal pathogenesis triggered by C. difficile toxin A.     

内生真菌Alternaria sp. GHX-P17代谢产物防治广藿香青枯病及保护酶的变化

广藿香是著名药材,青枯病是威胁广藿香生产和质量的主要病害。针对从广藿香茎叶中分离出的1株有抑菌活性的内生真菌Alternaria sp. GHX-P17,研究其代谢产物对广藿香青枯病的防治作用及耐病机制。在室内通过人工接种Alternaria sp. GHX-P17菌株与喷施粗提物稀释液2种方式,于不同时间调查广藿香青枯病的发病率和严重度,并计算其病情指数(disease index,DI)。同时,对处理后不同时间的广藿香苯丙氨酸解氨酶(phenylalanine ammonia lyase,PAL)、过氧化物酶(peroxidase,POD)和超氧化物歧化酶(superoxide dismutase,SOD)3种保护酶活性进行测定。结果表明:(1)Alternaria sp. GHX-P17处理后青枯病DI显著降低,与对照比较,204 h后DI降低为27.16%,方差分析呈显著性差异(P<0.05)。(2)随着处理时间延长,青枯病严重度升高缓慢,严重度等级降低,到204 h时,处理组的严重度明显低于对照,防治效果达到74.65%。(3)处理后的广藿香3种保护酶PAL、POD和SOD活性增强,但3种酶活性高峰出现时间不同。PAL随时间逐渐升高; POD先升高后降低,然后又升高,出现2个峰值; SOD快速升高后逐渐降低。这表明内生真菌菌株Alternaria sp. GHX-P17可以提高广藿香3种保护酶活性,延缓了青枯菌的侵染过程,降低了青枯病的发病程度。该研究结果为植物内生真菌次级代谢产物活性成分研究及生物农药开发提供了参考。     

Efficient generation of genome-modified mice via offset-nicking by CRISPR/Cas system

The mammalian zygote-mediated CRISPR/Cas system can efficiently generate targeted genome-modified animals. However, this system is limited by the risk of off-target mutations. Here we show that offset-nicking by Cas9 nickase and paired gRNAs allows us to generate region deleted mice and targeted knock-in mice without off-target mutations.     

A system for monitoring and improving animal visibility and its implications for zoological parks

With the growing trend in zoos to build complex, naturalistic exhibits comes the potential for exhibits to be so densely vegetated or complex that animals are not easily seen by zoo visitors. This can negatively impact the visitor's visiting experience and the zoo's ability to communicate conservation and education messages. Over the past 9 years, Disney's Animal Kingdom^® has developed a process for monitoring and improving the visibility of animals on display to the public. This animal visibility process utilizes a data collection system whereby systematic observations are collected each week. The percentage of observations where at least one animal was visible is recorded for each species and compared to an 80% visibility criterion. Species that do not reach this criterion for 4 consecutive weeks are discussed at animal management meetings. If the problems associated with animal visibility cannot be easily solved, the animal‐care teams partner with the research team to conduct a second process, called the Visibility Issues Process. This process provides additional information for the animal‐care team to utilize in developing a plan to improve visibility. Although the processes described here are specific to the infrastructure at Disney's Animal Kingdom^®, the basic concepts of (1) a formalized visibility data collection process, (2) a visibility criterion to which managers of species are held accountable, and (3) a process for planning to improve animal visibility without negatively impacting animal welfare are fundamental concepts that can be developed at individual institutions and incorporated into that zoo's existing infrastructure. Zoo Biol 29:68–79, 2010. © 2009 Wiley‐Liss, Inc.     

乳糖操纵子介导的半乳糖苷酶在转基因动物中的应用前景

细菌的乳糖操纵子可以在哺乳动物中调控基因的表达,修饰阻抑物基因和操纵基因可调控阻抑物诱导能力及其对操纵基因的亲和力,更好的适应高等动物的内环境。半乳糖苷酶对乳糖操纵子系统有正向调控作用,利用乳糖操纵子在转基因动物中可诱导性调控半乳糖苷酶基因的表达,能有效的提高转基因动物利用半乳糖苷、吸收营养物质的能力。以下从乳糖操纵子的结构功能、乳糖阻抑物功能活性的基因调控、操纵基因的功能以及其在哺乳动物的应用现状四个方面,结合半乳糖苷酶的生理功能和其在转基因动物中应用两个方面进行综述,对乳糖操纵子介导的半乳糖苷酶在转基因动物中的应用效果和前景进行了分析探讨。     

百日咳杆菌粘附素对猪源支气管败血波氏杆菌的保护性抗原的筛选

摘要：【目的】本研究通过百日咳杆菌黏附素(PRN)基因的分段克隆表达及其在BALB/c小鼠的主动和被动免疫保护试验筛选PRN中的保护性抗原肽。【方法和结果】利用大肠杆菌进行PRN的完整蛋白、N端和C端多肽及其RI和RII区域多肽（双拷贝）的表达,命名为GST-PRN、GST-PN、GST-PC、GST-2PRI和GST-2PRII。Western blot检测证实5种表达产物均具有良好的反应原性。在主动免疫保护试验中,5种表达产物均能诱导小鼠产生较高的PRN抗体水平;当使用3 LD50的支气管败血波氏杆菌     

猪丹毒丝菌天然SpaA和重组SpaA-N免疫保护效果的评价

【目的】本试验以小鼠为动物模型,评估了猪丹毒丝菌重组表面保护性抗原A的N端保护区域(rSpaA-N)和天然SpaA的免疫保护效果。【方法】将猪丹毒丝菌C43311株SpaA-N以可溶形式表达在大肠杆菌BL21中,用GST Bind Resin纯化试剂盒纯化rSpaA-N,采用电洗脱法从猪丹毒丝菌C43311株NaOH提取抗原中纯化天然SpaA,将rSpaA-N、天然SpaA和NaOH提取抗原制成亚单位疫苗,同时设GST及生理盐水对照组,间隔2周分3次皮下免疫小鼠,第3次免疫后2周用100LD50猪丹毒丝菌C43065株进行腹腔攻毒,采用间接ELISA方法检测免疫组小鼠血清的抗体动态变化。【结果】SDS-PAGE结果显示,采用GST Bind Resin纯化试剂盒和电洗脱法纯化得到了66kDa的rSpaA-N和64kDa的天然SpaA,蛋白含量分别为1.34mg/mL和1.26mg/mL,而Western印迹结果表明rSpaA-N和纯化前后的SpaA具有良好的免疫反应性。保护试验结果表明,不同免疫剂量的rSpaA-N组、天然SpaA组和NaOH提取抗原组均能完全保护小鼠受强毒株C43065的致死性攻击,而GST组和生理盐水组小鼠攻毒后全部死亡。ELISA检测结果表明,在不同免疫剂量的rSpaA-N组、天然SpaA组和NaOH提取抗原组小鼠血清中的抗体效价之间无显著差异(P0.05)。【结论】本研究结果表明rSpaA-N具有良好的免疫保护作用,可以作为猪丹毒亚单位疫苗。     

Recipient Environment More Important than Community Composition in Determining the Success of an Experimental Sponge Transplant

Human activities have inflicted profound damage upon many ecosystems, and ecologists are now seeking effective means of restoring ecosystems to their natural state. Industrial ports and harbors are highly modified and often depauperate in native fauna. They are typically characterized by poor water quality and modified community composition, both of which may hinder attempts to reintroduce native species. Here, we conducted a field experiment to separate the effects of the recipient environment and community composition on the success of endemic sponge explants in Port Kembla Harbor, NSW, Australia. A reciprocal transplant was conducted between communities originating from six sites that varied in water quality and community composition, enabling us to assess the relative factors simultaneously. A colony of the endemic sponge Tedania anhelans was then inserted into the center of each community, and we quantified the survival, growth, and metal bioaccumulation of sponges over three months. Endemic sponges consistently performed better against resident assemblages when water quality was good. Sponges transplanted to cleaner sites had over double the survivorship and approximately three times the surface area of sponges transplanted to disturbed sites. These patterns were independent of community composition. Bioaccumulation of metals in sponges was correlated with survival; however, other factors such as turbidity may be required to explain sponge mortality at some sites. This study adds to evidence that remediation of the physical and chemical environments may be a prerequisite for biological remediation and demonstrates the value of experimental transplants in assessing restoration potential.     

Livestock Predation—Insights From Problem Animal Control Registers in Botswana

Abstract: Problem Animal Control Registers, where farmers report livestock losses due to predators as a prerequisite for financial compensation, allow quantifying the human-predator conflict. We analyzed such registers from the Kweneng District of Botswana to assess the impact of native predators on livestock over 3 years. Leopards (Panthera pardus), lions (Panthera leo), wild dogs (Lycaon pictus), brown hyenas (Hyaena brunnea), and cheetahs (Acinonyx jubatus) altogether claimed 2,272 head of livestock. During 2002, the year with the highest impact, the number of animals reported depredated (954) represented 0.34% of the livestock in the district. Leopards and lions caused 64% of the losses. Leopard livestock predation mainly affected calves and was consistent over the district and over time. In contrast, lion predation concentrated on adult cattle, was characterized by local hot spots close to reserve borders, decreased with increasing distance to a reserve, and increased during 2002, an unusually dry year. Interviews with 60 farmers and herders within 30 km of Khutse and Central Kalahari Game Reserves revealed an annual loss of 2.2% of their livestock to predators. Here, small farms (max. 100 domestic animals) suffered relatively higher losses than large, commercial farms, not only due to predation (small farms: 11.7%; large farms: 1.0%) but also from other causes (small: 12.6%; large: 2.8%), even though herders on large farms guarded 5 times more livestock per person than those on small farms. To reduce livestock predation in most of the district where lions are absent, we recommend maternity corrals for pregnant females and calves to better protect vulnerable calves during day and night. In areas close to a reserve where lions roam, herders' incentives to keep all livestock protected in a corral at night have to be enhanced because, according to the registers, only 3 predation cases were reported to have happened inside a corral.